Construction and high cytoplasmic expression of a tumoricidal single-chain antibody against hepatocellular carcinoma |
| |
| Authors: | Duanpen Sandee Sumalee Tungpradabkul Manae Tsukio Tadayuki Imanaka Masahiro Takagi |
| |
| Affiliation: | (1) Department of Biotechnology, Graduate school of Engineering, Osaka University, Suita Osaka, 565-0871, Japan;(2) Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand;(3) Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan;(4) School of Materials Science, Japan Advanced Institute of Science and Technology, Ishikawa 923-1292, Japan |
| |
| Abstract: | ![]()
Background Hep27 monoclonal (Hep27 Mab) is an antibody against hepatocellular carcinoma. Hep27 Mab itself can inhibit the growth of a hepatocellular carcinoma cell line (HCC-S102). We attempted to produce a single-chain fragment (scFv), a small fragment containing an antigen-binding site of Hep27 Mab, by using DNA-recombinant techniques. Results The sequences encoding the variable regions of heavy (VH) and light (VL) chains of a murine Hep27 Mab were linked together by a linker peptide (Gly4Ser)3 and tagged with a hexa-histidine at the C-terminal; the resultant DNA construct was expressed in E. coli as an insoluble protein. The denatured scFv was refolded and purified by immobilized metal ion affinity chromatography (12 mg/l with a molecular weight of 27 kDa). Hep27scFv exhibited a tumoricidal activity against the HCC-S102 cell as its parental antibody (Hep27 Mab). Conclusion This scFv may be a potential candidate for a targeting agent in HCC immunodiagnosis or immunotherapy. |
| |
| Keywords: | |
| 本文献已被 SpringerLink 等数据库收录！ |
|
